Circulating HMGB1 is elevated in veterans with Gulf War Illness and triggers the persistent pro-inflammatory microglia phenotype in male C57Bl/6J mice

Gulf War Illness (GWI) is a chronic, multi-symptom peripheral and CNS condition with persistent microglial dysregulation, but the mechanisms driving the continuous neuroimmune pathology are poorly understood. The alarmin HMGB1 is an autocrine and paracrine pro-inflammatory signal, but the role of circulating HMGB1 in persistent neuroinflammation and GWI remains largely unknown. Using the LPS model of the persistent microglial pro-inflammatory response, male C57Bl/6J mice injected with LPS (5 mg/kg IP) exhibited persistent changes in microglia morphology and elevated pro-inflammatory markers in the hippocampus, cortex, and midbrain 7 days after LPS injection, while the peripheral immune response had resolved. Ex vivo serum analysis revealed an augmented pro-inflammatory response to LPS when microglia cells were cultured with the 7-day LPS serum, indicating the presence of bioactive circulating factors that prime the microglial pro-inflammatory response. Elevated circulating HMGB1 levels were identified in the mouse serum 7 days after LPS administration and in the serum of veterans with GWI. Tail vein injection of rHMGB1 in male C57Bl/6 J mice elevated TNFα mRNA levels in the liver, hippocampus, and cortex, demonstrating HMGB1-induced peripheral and CNS effects. Microglia isolated at 7 days after LPS injection revealed a unique transcriptional profile of 17 genes when compared to the acute 3 H LPS response, 6 of which were also upregulated in the midbrain by rHMGB1, highlighting a distinct signature of the persistent pro-inflammatory microglia phenotype. These findings indicate that circulating HMGB1 is elevated in GWI, regulates the microglial neuroimmune response, and drives chronic neuroinflammation that persists long after the initial instigating peripheral stimulus

Continuous Characterization of Universal Invertible Amplifier Using Source Noise

With passage of time and repeated usage of a system, component values that make up the system parameters change, causing errors in its functional output. In order to ensure the fidelity of the results derived from these systems it is thus very important to keep track of the system parameters while being used. This thesis introduces a method for tracking the existing system parameters while the system was being used using the inherent noise of its signal source. Kalman filter algorithm is used to track the inherent noise response to the system and use that response to estimate the system parameters. In this thesis this continuous characterization scheme has been used on a Universal Invertible Amplifier (UIA). Current biomedical research as well as diagnostic medicine depend a lot on shape profile of bio-electric signals of different sources, for example heart, muscle, nerve, brain etc. making it very important to capture the different event of these signals without the distortion usually introduced by the filtering of the amplifier system. The Universal Invertible Amplifier extracts the original signal in electrodes by inverting the filtered and compressed signal while its gain bandwidth profile allows it to capture from the entire bandwidth of bioelectric signals. For this inversion to be successful the captured compressed and filtered signals needs to be inverted with the actual system parameters that the system had during capturing the signals, not its original parameters. The continuous characterization scheme introduced in this thesis is aimed at knowing the system parameters of the UIA by tracking the response of its source noise and estimating its transfer function from that. Two types of source noises have been tried out in this method, an externally added noise that was digitally generated and a noise that inherently contaminates the signals the system is trying to capture. In our cases, the UIA was used to capture nerve activity from vagus nerve where the signal was contaminated with electrocardiogram signals providing us with a well-defined inherent noise whose response could be tracked with Kalman Filter and used to estimate the transfer function of UIA. The transfer function estimation using the externally added noise did not produce good results but could be improved by means that can be explored as future direction of this project. However continuous characterization using the inherent noise, a bioelectric signal, was successful producing transfer function estimates with minimal error. Thus this thesis was successful to introduce a novel approach for system characterization using bio-signal contamination

Inhalation Triggers Neuroimmune, Glial, and Neuropeptide Transcriptional Changes

Increasing evidence associates indoor fungal exposure with deleterious central nervous system (CNS) health, such as cognitive and emotional deficits in children and adults, but the specific mechanisms by which it might impact the brain are poorly understood. Mice were exposed to filtered air, heat-inactivated Aspergillus versicolo r (3 × 10 5 spores), or viable A. versicolo r (3 × 10 5 spores) via nose-only inhalation exposure 2 times per week for 1, 2, or 4 weeks. Analysis of cortex, midbrain, olfactory bulb, and cerebellum tissue from mice exposed to viable A. versicolo r spores for 1, 2, and 4 weeks revealed significantly elevated pro-inflammatory ( Tnf and Il1b ) and glial activity ( Gdnf and Cxc3r1 ) gene expression in several brain regions when compared to filtered air control, with the most consistent and pronounced neuroimmune response 48H following the 4-week exposure in the midbrain and frontal lobe. Bulk RNA-seq analysis of the midbrain tissue confirmed that 4 weeks of A. versicolo r exposure resulted in significant transcriptional enrichment of several biological pathways compared to the filtered air control, including neuroinflammation, glial cell activation, and regulation of postsynaptic organization. Upregulation of Drd1 , Penk , and Pdyn mRNA expression was confirmed in the 4-week A. versicolo r exposed midbrain tissue, highlighting that gene expression important for neurotransmission was affected by repeated A. versicolor inhalation exposure. Taken together, these findings indicate that the brain can detect and respond to A. versicolo r inhalation exposure with changes in neuroimmune and neurotransmission gene expression, providing much needed insight into how inhaled fungal exposures can affect CNS responses and regulate neuroimmune homeostasis